Geniposidic acid alleviates osteoarthritis progression through inhibiting inflammation and chondrocytes ferroptosis.

Osteoarthritis is one of the common diseases that seriously affects the quality of life of middle-aged and elderly people worldwide. Geniposidic acid (GPA) is extracted from Eucommia ulmoides that exhibits various pharmacological effects. This study investigated the function of GPA on osteoarthritis (OA) in IL-1ß-stimulated mouse chondrocytes and mouse OA model. Mouse OA model was established by destabilization of the medial meniscus (DMM) and GPA was given intraperitoneal injection. The results demonstrated that GPA could alleviate DMM-induced OA in mice. In vitro, IL-1ß-induced PGE2, NO, MMP1 and MMP3 were suppressed by GPA. Furthermore, IL-1ß-induced ferroptosis was inhibited by GPA, as confirmed by the inhibition of MDA, iron, and ROS, as well as the upregulation of GSH, GPX4, and Ferritin. In addition, GPA was found to increase the expression of Nrf2 and HO-1. And the inhibition of GPA on IL-1ß-induced inflammation and ferroptosis were prevented by Nrf2 inhibitor. In conclusion, GPA alleviates OA progression through inhibiting inflammation and chondrocytes ferroptosis via Nrf2 signalling pathway.

Exosome microRNA-22 inhibiting proliferation, migration and invasion through regulating Twist1/CADM1 axis in osteosarcoma.

This study aims to the function of miR-22 original mesenchymal stem cells (MSC) on osteosarcoma (OS) proliferation, migration and invasion. Bio-informatics analysis including GEO2R analysis, Gene Ontology analysis, integration analysis were used to confirmed the target genes (miR-22, Twist1, CADM1) in OS. RT-qPCR and western blotting confirmed the different expression of miR-22, Twist1, CADM1 in OS tissues, MG63 and Saos cell lines. MTS assay, CCK8 assay, colony forming assay, EdU assay were performed to detect the proliferation effect of miR-22 on MG63. Transwell migration assay, transwell invasion assay, wound healing assay were used to verify the migration and invasion effect of miR-22 on MG63. Luciferase reporter assay confirm the binding sites between miR-22 and Twist1. RT-qPCR confirmed miR-22 and CADM1 downregulated and Twist1 upregulated in OS tissues, MG63 and Saos. Exosome original MSC labeled with PKH-26 could be uptake by MG63, which upregulated the expression of miR-22 in MG63. High expression of miR-22 in MG63 inhibited proliferation, migration and invasion, which could be rescued by Twist1. Dual luciferase reporter analysis confirmed Twist1 was a target of miR-22. Exosome modified with miR-22 mimic inhibit proliferation, migration and invasion more efficient than exosome original MSC. miR-22 cargo in exo-MSC could uptake by MG63 and supply MG63 with miR-22, which inhibit MG63 proliferation, migration and invasion through targeting Twist1.

Ruscogenin attenuates cartilage destruction in osteoarthritis through suppressing chondrocyte ferroptosis via Nrf2/SLC7A11/GPX4 signaling pathway.

Osteoarthritis (OA) is a common joint degenerative disease, and chondrocyte injury is the main pathological and physiological change. Ruscogenin (Rus), a bioactive compound isolated from Radix Ophiopogon japonicus, exhibits various pharmacological effects. The aim of this research was to test the role and mechanism of Rus on OA both in vivo and in vitro. Destabilized medial meniscus (DMM)-induced OA model was established in vivo and IL-1ß-stimulated mouse chondrocytes was used to explore the role of Rus on OA in vitro. In vivo, Rus exhibited protective effects against DMM-induced OA model. Rus could inhibit MMP1 and MMP3 expression in OA mice. In vitro, IL-1ß-induced inflammation and degradation of extracellular matrix were inhibited by Rus, as confirmed by the inhibition of PGE2, NO, MMP1, and MMP3 by Rus. Also, IL-1ß-induced ferroptosis was suppressed by Rus, as confirmed by the inhibition of MDA, iron, and ROS, as well as the upregulation of GSH, GPX4, Ferritin, Nrf2, and SLC7A11 expression induced by Rus. Furthermore, the suppression of Rus on IL-1ß-induced inflammation, MMPs production, and ferroptosis were reversed when Nrf2 was knockdown. In conclusion, Rus attenuated OA progression through inhibiting chondrocyte ferroptosis via Nrf2/SLC7A11/GPX4 signaling pathway.

Brevilin A attenuates cartilage destruction in osteoarthritis mouse model by inhibiting inflammation and ferroptosis via SIRT1/Nrf2/GPX4 signaling pathway.

Osteoarthritis (OA) is a serious orthopedic disease that affects people's quality of life. Although there are many treatment methods, the treatment effect is still not good. Brevilin A is a bioactive compound isolated from the medicinal herbCentipeda minima. The potential efficacy of brevilin A on OA was explored in this study. Mouse chondrocytes were isolated and stimulated by IL-1ß and mouse OA model was induced by destabilization of the medial meniscus (DMM). The results demonstrated that brevilin A markedly inhibited IL-1ß-induced MMP1 and MMP3 production. IL-1ß-induced PGE2, NO, MDA, and iron production were alleviated by brevilin A. The production of GSH and the expression of SIRT1, Nrf2, HO-1, GPX4, and Ferritin were increased by brevilin A. Furthermore, the inhibition of brevilin A on IL-1ß-induced inflammation and ferroptosis were prevented by SIRT1 inhibitor. In vivo, the results showed brevilin A markedly attenuated OA progression in DMM-induced mouse OA model. Also, brevilin A could alleviate MMP1, MMP3, iNOS, and COX2 expression in OA mice. In conclusion, brevilin A protected mice against OA via suppressing inflammatory response and ferroptosis by regulating SIRT1/Nrf2/GPX4 signaling.

Kukoamine A protects mice against osteoarthritis by inhibiting chondrocyte inflammation and ferroptosis via SIRT1/GPX4 signaling pathway.

AIMS: Osteoarthritis (OA) is one of the common chronic degenerative joint diseases, characterized by cartilage damage, subchondral bone changes, osteophyte formation, and synovitis. Kukoamine A (KuKA) is a bioactive compound isolated from Lycium chinense which is known as its anti-inflammatory activity. In this study, we detected the regulatory role of KuKA on OA both in vivo and in vitro. MATERIALS AND METHODS: Mouse chondrocytes were cultured and mouse model of OA was established. Inflammatory mediator was measured by ELISA. The signaling pathway was tested by western blot analysis. KEY FINDINGS: KuKA inhibited IL-1ß-induced PGE2 and NO production and iNOS and COX-2 expression. IL-1ß-induced MMP1 and MMP3 production was attenuated by KuKA. IL-1ß-induced MDA, iron, and ROS were alleviated by KuKA. Meanwhile, GSH content, GPX4, Ferritin, SIRT1, Nrf2, and HO-1 expression were upregulated by KuKA. Furthermore, the inhibitory role of KuKA on IL-1ß-induced inflammation, MMPs production, and ferroptosis were reversed by SIRT1 inhibitor. In vivo, KuKA could attenuate OA development in mouse model. KuKA markedly alleviated MMP1, MMP3, iNOS, and COX2 expression in OA mice. SIGNIFICANCE: In conclusion, KuKA could inhibit OA development through suppressing chondrocyte inflammation and ferroptosis via SIRT1/GPX4 signaling pathway.

circAGFG1 sponges miR-28-5p to promote non-small-cell lung cancer progression through modulating HIF-1α level.

Circular RNAs (circRNAs) have gained much attention for their crucial regulatory roles in human diseases and cancers. However, the role and the mechanism of circRNA ArfGAP with FG repeats 1 (circAGFG1) in non-small-cell lung cancer (NSCLC) are still largely unknown. circAGFG1 was highly expressed in NSCLC, and high expression of circAGFG1 was closely related to the low survival rate of NSCLC patients. circAGFG1 knockdown inhibited the proliferation, migration, and invasion and promoted the apoptosis of NSCLC cells. circAGFG1 bound to miR-28-5p in NSCLC cells, and circAGFG1 promoted NSCLC progression partly through sponging miR-28-5p in vitro. HIF-1α was a target of miR-28-5p, and miR-28-5p overexpression-mediated influences in NSCLC cells were partly overturned by the addition of HIF-1α overexpression plasmid. circAGFG1/miR-28-5p/HIF-1α axis regulated cellular glycolytic metabolism in NSCLC cells. circAGFG1 silencing restrained the xenograft tumor growth in vivo. circAGFG1 promoted the proliferation, migration, and invasion and suppressed the apoptosis of NSCLC cells through accelerating the glycolysis via miR-28-5p/HIF-1α axis.

Controlled fabrication of core-shell silica@chiral metal-organic framework for significant improvement chromatographic separation of enantiomers.

Chiral metal-organic frameworks (CMOFs) have been explored as potential chiral stationary phases (CSPs) for chiral high performance liquid chromatography (HPLC). However, their application is still hindered by the low column efficiency, high back pressure and the difficulty in column packing due to the irregular shapes and wide size distributions of CMOF particles. Here we report an efficient one-pot method for the immobilization of chiral MOF [Cu2((+)-Cam)2Dabco] (Cu2C2D) onto microspheric silica particles, generating a uniform core-shell microsphere with SiO2@CMOF@CMOF morphology as CSP packing material. Significantly, the shell thickness and the corresponding column efficiency could be rationally regulated by controlling the growth cycles of [Cu2((+)-Cam)2Dabco]. The structure of developed core-shell microspheres were characterized by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), powder X-ray diffraction (PXRD), N2 adsorption experiments, infrared spectroscopy (FT-IR) and thermogravimetric analysis. Mechanism involved in the chromatographic separation is the multi-interactions including hydrophobic and hydrogen-bonding, etc. Based on these interactions, successful separation could be achieved among different types of racemic compounds such as carboxylic acid, ketones and phenols under normal phase liquid chromatography (NPLC) condition. In addition, the relative standard deviation (RSD%) value of the retention time for TNPTO was below 1.1% (n = 5), indicating the good repeatability and stability of the chiral SiO2@Cu2C2D-2 column for HPLC enantioseparation. The results reveal that the CMOF coating approach is convenient to fabricate CSP with pre-designed functions of good recognition performance and to facilitate the evolution of CSP in chiral HPLC. Furthermore, the SiO2@Cu2C2D-2 column was successfully employed for the determination of the enantiomeric excess value for TNPTO in the asymmetric Michael addition reaction.

[Synthesis of a chiral metal-organic framework using zinc oxide as zinc source for enantioselective adsorption of 2,2'-furoin racemate].

Chiral metal-organic frameworks (CMOFs),[Zn(L-mal)(H2O)2]n were prepared by using zinc oxide instead of the traditional metal inorganic salt as the zinc source, and their chiral separation performance was investigated. Equal amounts of ZnO and L-malic acid were dissolved in H2O, and[Zn(L-mal)(H2O)2]n were obtained by allowing the mixture to stand at room temperature for 24 h. Then, 100 mg[Zn(L-mal)(H2O)2]n as the adsorbent was added to 250 µL 1 g/L 2,2'-furoin racemate, and 4.5 mL isopropanol and 1 mL methanol were selected as the extraction solvent and elution solvent, respectively, for selective adsorption experiments. The final filtrate was analyzed by high performance liquid chromatography (HPLC). The results showed that[Zn(L-mal)(H2O)2]n exhibited better selective adsorption ability for R-2,2'-furoin with 20% enantiomeric excess (ee). This work provides a new method for the green preparation of CMOF and expands its application in the field of chiral separation.

Revelation of the chiral recognition of alanine and leucine in an l-phenylalanine-based metal-organic framework.

As a proof-of-concept, an l-phenylalanine-based metal-organic framework Zn-MOF, namely [Zn2(l-Phe)2(bpe)2]n, was designed for experimentally revealing the chiral recognition mechanism of alanine and leucine by means of 13C CP MAS NMR spectroscopy, X-ray photoelectron spectroscopy and control experiments.

Tetramethylpyrazine attenuates periorbital allodynia and neuroinflammation in a model of traumatic brain injury.

BACKGROUND: Traumatic brain injury (TBI) is a public health issue. As the major complaint in 51% of TBI patients, chronic pain is an important aspect in TBI treatment. Tetramethylpyrazine (TMP) is an important compound in Ligustrazine, an analgesic drug in traditional Chinese medicine, but its potential in relieving pain symptom in TBI has not been tested. We established a TBI mouse model with controlled cortical impact (CCI), and measured periorbital hypersensitivity with von Frey monofilaments. We examined activated microglia and astrocytes and the levels of substance P (SP) and inducible isoform of nitric oxide synthase (iNOS) with immunohistochemistry, measured mRNA and protein levels of proinflammatory cytokines with qPCR and enzyme-linked immunosorbent assay, respectively. Western blot was employed to detect molecules in NF-κB signaling pathway. RESULTS: TMP significantly attenuated periorbital hypersensitivity in TBI mice. Within 3 days after CCI, TMP attenuated activation of microglia and astrocytes, levels of SP, iNOS, and CGRP in trigeminal pathway, and levels of proinflammatory cytokines (including IL-6, TNF-α, IL-12). In isolated microglia, TMP attenuated the effects of lipopolysaccharide on the phosphorylation of cytoplasmic IKKα/ß and IKB-α, and levels of nucleic p65. CONCLUSION: TMP reversed periorbital hypersensitivity by limiting neuroinflammation at the primary stage of TBI, and could be a promising drug for pain treatment in TBI.

[Correlation of Treg and IL-15 expression in the peripheral blood of patients with oral lichen planus].

PURPOSE: To investigate the expression of Treg and IL-15 in the peripheral blood of patients with oral lichen planus (OLP). METHODS: The peripheral blood was obtained from 36 OLP patients and 20 healthy controls. Flow cytometry was used to evaluate the proportion of Treg cells, and the level of IL-15 was measured by ELISA. The data were analyzed by SPSS 16.0 software package. RESULTS: In the peripheral blood of OLP patients, the proportion of CD4+CD25+Foxp3+ Treg cells and IL-15 content were significantly higher than those of the healthy controls (P<0.05), but there were no significantly difference between two clinical subtypes of OLP(P>0.05). The proportion of CD4+CD25+Foxp3+Treg cells was positively correlated with the level of IL-15(r=0.70,P<0.05). CONCLUSIONS: The high expression of Treg cells and IL-15 in the perpheral blood of OLP patients may be responsible for the pathogenesis of OLP.

[Purification of recombinant human antithrombin III expressed in a goat mammary bioreactor].

Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.